Phosphoramide compounds

ABSTRACT

The invention provides a compound of formula I:  
                 
 
     wherein R 1 , R a , R b , R c , and R d  have any of the values defined in the specification, as well as pharmaceutical compositions comprising such compounds or salts. The compounds are useful for treating cancer in animals.

RELATED APPLICATIONS

[0001] This application is a continuation under 37 C.F.R. 1.53(b) ofU.S. application Ser. No. 10/047,465 filed Jan. 14, 2002, which is acontinuation under 35 USC 111 (a) of International Application No.PCT/US00/19361 filed Jul. 14, 2000 and published in English as WO01/04130 A1 on Jan. 18, 2001, which claims priority from U.S.Provisional Application Serial No. 60/143,799 filed Jul. 14, 1999, whichapplications and publication are incorporated herein by reference.

UNITED STATES GOVERNMENT FUNDING

[0002] The invention described herein was made with government supportunder Grant Number CA34619 awarded by the National Cancer Institute, andunder Grant Number GM08298 awarded by the National Institutes ofHealth—National Institute of General Medical Sciences PredoctoralTraining Grant in Chemical Pharmacology. The United States Governmenthas certain rights in the invention.

BACKGROUND OF THE INVENTION

[0003] Increased selectivity of anti-cancer agents is an importantfactor in designing new drugs for the treatment of cancer. The designand synthesis of novel compounds that can be activated selectively incancer cells is therefore an attractive way to target the inhibition oftumor growth. This strategy ensures that cytotoxicity occurs selectivelyin malignant cells and might lead to a reduction of many of the sideeffects commonly caused by chemotherapeutic agents currently in use. Anapproach to the enhancement of selectivity for cytotoxic chemotherapyinvolves the design of prodrugs that undergo preferential activation byenzymes that are overexpressed in tumors. These prodrugs, which are notcytotoxic until they are metabolically activated, can serve to deliverselectively the cytotoxic agent to the tumor site.

[0004] One such prodrug that is used clinically is the compoundcyclophosphamide (1, FIG. 1). Cyclophosphamide is activated to4-hydroxycyclophosphamide (1a) by hepatic cytochrome P-450 oxidation(FIG. 1). Subsequent β-elimination from the aldehyde tautomer of 1areleases phosphoramide mustard 2 as the active drug which can cyclizeintramolecularly to a short-lived electrophilic aziridinium ionintermediate (3). Nucleophilic addition can then occur and thecyclization/addition process can be repeated. Ultimately, if DNA is thenucleophile, the phosphoramide mustard can cross-link DNA and inhibitfurther DNA replication, a process that leads to cell death.

[0005] The design of chemotherapeutic quinone prodrugs that arebioreductively activated by the enzyme DT-diaphorase has also beeninvestigated (see for example P. Workman, Oncol. Res., 1994, 6,461-475;and R. J. Riley and P. Workman, Biochem. Pharmacol., 1992, 43,1657-1669). Many of the compounds studied include benzimidazolequinone,benzoquinone and naphthoquinone prodrugs, with and without an alkylatingmoiety attached to the core ring structure, and indolequinone analogspatterned after the known cytotoxic agents Mitomycin C and E09.Sartorelli et al prepared a series of naphthoquinone prodrugs that couldpotentially be transformed into alkylating moieties following theexpulsion of a leaving group from the bioreductively activated compound(see for example, N. E. Sladek, Pharmac. Ther., 1988, 37, 301-355; andM. Colvin et al., Cancer Res., 1976, 36, 1121-1126).

[0006] Despite the reported success in treating cancer with the compoundCyclophosphamide, there is currently a need for structurally noveltherapeutic agents that can be used to treat cancer.

SUMMARY OF THE INVENTION

[0007] Applicant has discovered a series of novel compounds that possessuseful cytotoxic properties when administered in vivo. Accordingly, theinvention provides a compound of formula I:

[0008] wherein:

[0009] R¹ is an organic releasing group comprising a quinone ring;

[0010] R_(a), R_(b), R_(c), and R_(d) are each independently hydrogen,(C₁-C₆)alkyl, or —CH₂CH₂X; and

[0011] each X is independently halo, (C₁-C₆)alkylsulfonyl,halo(C₁-C₆)alkylsulfonyl, or arylsulfonyl, wherein each aryl isoptionally substituted with one or more (e.g. 1, 2, 3, or 4) halo,(C₁-C₆)alkyl, halo(C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)alkanoyl,(C₁-C₆)alkanoyloxy, (C₁-C₆)alkoxycarbonyl, cyano, nitro, ortrifluoromethoxy;

[0012] provided at least two of R_(a), R_(b), R_(c), and R_(d) are—CH₂CH₂X;

[0013] or a pharmaceutically acceptable salt thereof.

[0014] The invention also provides a pharmaceutical compositioncomprising a compound of formula I, or a pharmaceutically acceptablesalt thereof, in combination with a pharmaceutically acceptable diluentor carrier.

[0015] Additionally, the invention provides a therapeutic method forpreventing or treating cancer (e.g. a tumor) comprising administering toa mammal in need of such therapy, an effective amount of a compound offormula I, or a pharmaceutically acceptable salt thereof.

[0016] The invention also provides a compound of formula I for use inmedical therapy (preferably for use in treating cancer, such as a tumor)as well as the use of a compound of formula I for the manufacture of amedicament for the treatment of cancer (e.g. a tumor) in a mammal, suchas a human.

[0017] The invention also provides processes and intermediates usefulfor preparing compounds of formula I, or salts thereof.

BRIEF DESCRIPTION OF THE FIGURES

[0018]FIG. 1 illustrates the activation of cyclophosphamide in vivo.

DETAILED DESCRIPTION

[0019] The following definitions are used, unless otherwise described:halo is fluoro, chloro, bromo, or iodo. Alkyl, alkoxy, etc., denote bothstraight and branched groups; but reference to an individual radicalsuch as “propyl” embraces only the straight chain radical, a branchedchain isomer such as “isopropyl” being specifically referred to. Aryldenotes a phenyl radical or an ortho-fused bicyclic carbocyclic radicalhaving about nine to ten ring atoms in which at least one ring isaromatic.

[0020] As used herein the term “organic releasing group comprising aquinone ring” includes mono-, bi- and poly-cyclic ring systems thatcomprise at least one quinone ring, which ring systems are capable ofreleasing a group of formula (V):

[0021] from a compound of formula I when the compound of formula I isadministered to a mammal (e.g. a human). Preferred releasing groupsinclude cyclic ring systems that can be reduced in vitro by the enzymeDT-diaphorase, leading to the release of the group of formula (V). Otherpreferred releasing groups include cyclic ring systems that can besubstituted in vitro by glutathione (or other nucleophyles), leading tothe release of the group of formula (V). A more preferred releasinggroups is a group of formula II, III, or IV as described herein.

[0022] As used herein the term preventing or treating cancer includeskilling cancer cells and/or inhibiting their growth or proliferation.

[0023] It will be appreciated by those skilled in the art that compoundsof the invention having a chiral center may exist in and be isolated inoptically active and racemic forms. Some compounds may exhibitpolymorphism. It is to be understood that the present inventionencompasses any racemic, optically-active, polymorphic, orstereoisomeric form, or mixtures thereof, of a compound of theinvention, which possess the useful properties described herein, itbeing well known in the art how to prepare optically active forms (forexample, by resolution of the racemic form by recrystallizationtechniques, by synthesis from optically-active starting materials, bychiral synthesis, or by chromatographic separation using a chiralstationary phase) and how to determine anti-cancer activity using thestandard tests described herein, or using other similar tests which arewell known in the art.

[0024] Specific and preferred values listed below for radicals,substituents, and ranges, are for illustration only; they do not excludeother defined values or other values within defined ranges for theradicals and substituents.

[0025] Specifically, (C₁-C₆)alkyl can be methyl, ethyl, propyl,isopropyl, butyl, iso-butyl, sec-butyl, pentyl, 3-pentyl, or hexyl;(C₁-C₆)alkoxy can be methoxy, ethoxy, propoxy, isopropoxy, butoxy,iso-butoxy, sec-butoxy, pentoxy, 3-pentoxy, or hexyloxy; (C₁-C₆)alkanoylcan be acetyl, propanoyl or butanoyl; halo(C₁-C₆)alkyl can beiodomethyl, bromomethyl, chloromethyl, fluoromethyl, trifluoromethyl,2-chloroethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, or pentafluoroethyl;(C₁-C₆)alkoxycarbonyl can be methoxycarbonyl, ethoxycarbonyl,propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, pentoxycarbonyl, orhexyloxycarbonyl; (C₁-C₆)alkylthio can be methylthio, ethylthio,propylthio, isopropylthio, butylthio, isobutylthio, pentylthio, orhexylthio; and (C₁-C₆)alkanoyloxy can be formyloxy, acetoxy,propanoyloxy, butanoyloxy, isobutanoyloxy, pentanoyloxy, or hexanoyloxy.

[0026] A specific value for R¹ is a group of the formula (II):

[0027] wherein Re is hydrogen, halo, (C₁-C₆)alkyl, halo(C₁-C₆)alkyl,(C₁-C₆)alkoxy, (C₁-C₆)alkanoyloxy, cyano, nitro, or (C₁-C₆)alkylthio;and wherein the benz ring is optionally substituted by one or more (e.g.1, 2, 3, or 4) hydroxy, halo, (C₁-C₆)alkyl, halo(C₁-C₆)alkyl,(C₁-C₆)alkoxy, (C₁-C₆)alkylthio; (C₁-C₆)alkanoyl, (C₁-C₆)alkanoyloxy,(C₁-C₆)alkoxycarbonyl, cyano, nitro, mercapto, trifluoromethoxy, orNR_(f)R_(g); wherein each R_(f) and R_(g) is independently hydrogen,(C₁-C₆)alkyl, (C₁-C₆)alkanoyl, phenyl, benzyl, or phenethyl; or R_(f)and R_(g) together with the nitrogen to which they are attached arepyrrolidino, piperidino or morpholino.

[0028] Another specific value for R¹ is a group of the formula (III):

[0029] wherein R_(k) is hydrogen or (C₁-C₆)alkyl; R_(m) is hydrogen or(C₁-C₆)alkyl, phenyl, benzyl, or phenethyl; and wherein the benz ring isoptionally substituted by one or two hydroxy, halo, (C₁-C₆)alkyl,halo(C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)alkylthio; (C₁-C₆)alkanoyl,(C₁-C₆)alkanoyloxy, (C₁-C₆)alkoxycarbonyl, cyano, nitro, mercapto,trifluoromethoxy, or NR_(f)R_(g); wherein each R_(f) and R_(g) isindependently hydrogen, (C₁-C₆)alkyl, (C₁-C₆)alkanoyl, phenyl, benzyl,or phenethyl; or R_(f) and R_(g) together with the nitrogen to whichthey are attached are pyrrolidino, piperidino or morpholino.

[0030] Another specific value for R¹ is a group of the formula (IV):

[0031] wherein R_(n) is hydrogen or (C₁-C₆)alkyl; R_(p) is hydrogen or(C₁-C₆)alkyl, phenyl, benzyl, or phenethyl; and wherein the benz ring isoptionally substituted by one or two hydroxy, halo, (C₁-C₆)alkyl,halo(C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)alkylthio; (C₁-C₆)alkanoyl,(C₁-C₆)alkanoyloxy, (C₁-C₆)alkoxycarbonyl, cyano, nitro, mercapto,trifluoromethoxy, or NR_(f)R_(g); wherein each R_(f) and R_(g) isindependently hydrogen, (C₁-C₆)alkyl, (C₁-C₆)alkanoyl, phenyl, benzyl,or phenethyl; or R_(f) and R_(g) together with the nitrogen to whichthey are attached are pyrrolidino, piperidino or morpholino.

[0032] A specific value for X is bromo, chloro, mesyl,trifluoromethylsulfonyl, or tosyl. A more specific value for X is bromo.

[0033] A specific value for R_(e) is hydrogen, halo, methyl, ormethylthio.

[0034] A specific value for R_(h) is hydrogen or methyl.

[0035] A specific value for R_(k) is hydrogen or methyl.

[0036] A specific value for R_(m) is hydrogen or methyl.

[0037] A specific value for R_(n) is hydrogen or methyl.

[0038] A specific value for R_(p) is hydrogen or methyl.

[0039] A specific group of compounds of formula I are compounds whereinR_(a) is (C₁-C₆)alkyl.

[0040] A specific group of compounds of formula I are compounds whereinR_(c) is (C₁-C₆)alkyl.

[0041] A specific group of compounds of formula I are compounds whereinR_(a) and R_(b) are each independently —CH₂CH₂X.

[0042] A specific group of compounds of formula I are compounds whereinR_(c), and Rd are each independently —CH₂CH₂X.

[0043] A specific group of compounds of formula I are compounds whereinR_(b) and R_(d) are each independently —CH₂CH₂X.

[0044] A more specific group of compounds of formula I are compoundswherein R_(a) is methyl.

[0045] A more specific group of compounds of formula I are compoundswherein R_(c) is methyl.

[0046] A more specific group of compounds of formula I are compoundswherein R_(a) and R_(b) are each independently —CH₂CH₂Br.

[0047] A more specific group of compounds of formula I are compoundswherein R_(c), and R_(d) are each independently —CH₂CH₂Br.

[0048] A more specific group of compounds of formula I are compoundswherein R_(b) and R_(d) are each independently —CH₂CH₂Br.

[0049] A more specific group of compounds of formula I are compoundswherein R_(a) and R_(b) are each independently —CH₂CH₂Cl.

[0050] A more specific group of compounds of formula I are compoundswherein R_(c), and R_(d) are each independently —CH₂CH₂Cl.

[0051] A more specific group of compounds of formula I are compoundswherein R_(b) and R_(d) are each independently —CH₂CH₂Cl.

[0052] A preferred compound of the invention is:

[0053] 2-(1,4-naphthoquinonyl)methyl N,N-bis(2-chloroethyl)phosphorodiamidate;

[0054] 2-(3-Methyl-1,4-naphthoquinonyl)methyl N,N-bis(2-chloroethyl)phosphorodiamidate;

[0055] 2-(3-Thiomethyl-1,4-naphthoquinonyl)methyl N,N-bis(2-chloroethyl)phosphorodiamidate;

[0056] 2-(3-Bromo-1,4-naphthoquinonyl)methyl N,N-bis(2-chloroethyl)phosphorodiamidate;

[0057] 2-(1,4-Naphthoquinonyl)methyl N,N-bis(2-bromoethyl)phosphorodiamidate;

[0058] 2-(3-Methyl-1,4-naphthoquinonyl)methyl N,N-bis(2-bromoethyl)phosphorodiamidate;

[0059] 2-(1,4-Naphthoquinonyl)methyl bis[N-(2-chloroethyl)]phosphorodiamidate;

[0060] 2-(1,4-Naphthoquinonyl)methylbis[N-methyl-N-(2-bromoethyl)]phosphorodiamidate;

[0061] 2-(3-Methyl-1,4-naphthoquinonyl)methylbis[N-methyl-N-(2-bromoethyl)] phosphorodiamidate;

[0062] 2-(1,4-Naphthoquinonyl)methyl bis[N-methyl-N-(2-chloroethyl)]phosphorodiamidate;

[0063] 3-(5-Methoxy-1-methyl-4,7-indolequinonyl)-methylbis[N-methyl-N-(2-bromoethyl)] phosphorodiamidate;

[0064] 3-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylN,N-bis(2-bromoethyl)-phosphorodiamidate;

[0065] 2-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylbis[N-methyl-N-(2-bromoethyl)]phosphorodiamidate;

[0066] 2-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylN,N-bis(2-chloroethyl)-phosphorodiamidate; or

[0067] 2-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylN,N-bis(2-bromoethyl)-phosphorodiamidate;

[0068] or a pharmaceutically acceptable salt thereof.

[0069] A more prefered compound of the invention is:

[0070] 3-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylN,N-bis(2-bromoethyl)-phosphorodiamidate;

[0071] 2-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylbis[N-methyl-N-(2-bromoethyl)]phosphorodiamidate;

[0072] 2-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylN,N-bis(2-chloroethyl)-phosphorodiamidate; or

[0073] 2-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylN,N-bis(2-bromoethyl)-phosphorodiamidate;

[0074] or a pharmaceutically acceptable salt thereof.

[0075] Processes for preparing compounds of formula I are provided asfurther embodiments of the invention and are illustrated by thefollowing procedures in which the meanings of the generic radicals areas given above unless otherwise qualified.

[0076] A compound of formula I wherein R¹ is a group of formula II canbe prepared by oxidizing a corresponding compound wherein R¹ is a groupof formula VI:

[0077] wherein each R is independently (C₁-C₆)alkyl. The oxidation canbe carried out under standard conditions, for example using cericammonium nitrate. Fremy's salt, or silver (II) oxide. Suitableconditions for carrying out such an oxidation are described in theExamples hereinbelow.

[0078] A compound of formula I wherein R¹ is a group of formula III canbe prepared by oxidizing a corresponding compound wherein R¹ is a groupof formula VII:

[0079] wherein each R is independently (C₁-C₆)alkyl. The oxidation canbe carried out under standard conditions, for example using cericammonium nitrate, Fremy's salt, or silver (II) oxide. Suitableconditions for carrying out such an oxidation are described in theExamples hereinbelow.

[0080] A compound of formula I wherein R¹ is a group of formula IV canbe prepared by oxidizing a corresponding compound wherein R¹ is a groupof formula VIII:

[0081] wherein each R is independently (C₁-C₆)alkyl. The oxidation canbe carried out under standard conditions, for example using cericammonium nitrate, Fremy's salt, or silver (II) oxide. Suitableconditions for carrying out such an oxidation are described in theExamples hereinbelow.

[0082] An intermediate useful for preparing a compound of formula I is acorresponding compound of formula I wherein R¹ is a group of formula VI,VII, or VIII.

[0083] In cases where compounds are sufficiently basic or acidic to formstable nontoxic acid or base salts, administration of the compounds assalts may be appropriate. Examples of pharmaceutically acceptable saltsare organic acid addition salts formed with acids which form aphysiological acceptable anion, for example, tosylate, methanesulfonate,acetate, citrate, malonate, tartrate, succinate, benzoate, ascorbate,α-ketoglutarate, and α-glycerophosphate. Suitable inorganic salts mayalso be formed, including hydrochloride, sulfate, nitrate, bicarbonate,and carbonate salts.

[0084] Pharmaceutically acceptable salts may be obtained using standardprocedures well known in the art, for example by reacting a sufficientlybasic compound such as an amine with a suitable acid affording aphysiologically acceptable anion. Alkali metal (for example, sodium,potassium or lithium) or alkaline earth metal (for example calcium)salts of carboxylic acids can also be made.

[0085] The compounds of formula I can be formulated as pharmaceuticalcompositions and administered to a mammalian host, such as a humanpatient in a variety of forms adapted to the chosen route ofadministration, i.e., orally or parenterally, by intravenous,intramuscular, topical or subcutaneous routes.

[0086] Thus, the present compounds may be systemically administered,e.g., orally, in combination with a pharmaceutically acceptable vehiclesuch as an inert diluent or an assimilable edible carrier. They may beenclosed in hard or soft shell gelatin capsules, may be compressed intotablets, or may be incorporated directly with the food of the patient'sdiet. For oral therapeutic administration, the active compound may becombined with one or more excipients and used in the form of ingestibletablets, buccal tablets, troches, capsules, elixirs, suspensions,syrups, wafers, and the like. Such compositions and preparations shouldcontain at least 0.1% of active compound. The percentage of thecompositions and preparations may, of course, be varied and mayconveniently be between about 2 to about 60% of the weight of a givenunit dosage form. The amount of active compound in such therapeuticallyuseful compositions is such that an effective dosage level will beobtained.

[0087] The tablets, troches, pills, capsules, and the like may alsocontain the following: binders such as gum tragacanth, acacia, cornstarch or gelatin; excipients such as dicalcium phosphate; adisintegrating agent such as corn starch, potato starch, alginic acidand the like; a lubricant such as magnesium stearate; and a sweeteningagent such as sucrose, fructose, lactose or aspartame or a flavoringagent such as peppermint, oil of wintergreen, or cherry flavoring may beadded. When the unit dosage form is a capsule, it may contain, inaddition to materials of the above type, a liquid carrier, such as avegetable oil or a polyethylene glycol. Various other materials may bepresent as coatings or to otherwise modify the physical form of thesolid unit dosage form. For instance, tablets, pills, or capsules may becoated with gelatin, wax, shellac or sugar and the like. A syrup orelixir may contain the active compound, sucrose or fructose as asweetening agent, methyl and propylparabens as preservatives, a dye andflavoring such as cherry or orange flavor. Of course, any material usedin preparing any unit dosage form should be pharmaceutically acceptableand substantially non-toxic in the amounts employed. In addition, theactive compound may be incorporated into sustained-release preparationsand devices.

[0088] The active compound may also be administered intravenously orintraperitoneally by infusion or injection. Solutions of the activecompound or its salts can be prepared in water, optionally mixed with anontoxic surfactant. Dispersions can also be prepared in glycerol,liquid polyethylene glycols, triacetin, and mixtures thereof and inoils. Under ordinary conditions of storage and use, these preparationscontain a preservative to prevent the growth of microorganisms.

[0089] The pharmaceutical dosage forms suitable for injection orinfusion can include sterile aqueous solutions or dispersions or sterilepowders comprising the active ingredient which are adapted for theextemporaneous preparation of sterile injectable or infusible solutionsor dispersions, optionally encapsulated in liposomes. In all cases, theultimate dosage form should be sterile, fluid and stable under theconditions of manufacture and storage. The liquid carrier or vehicle canbe a solvent or liquid dispersion medium comprising, for example, water,ethanol, a polyol (for example, glycerol, propylene glycol, liquidpolyethylene glycols, and the like), vegetable oils, nontoxic glycerylesters, and suitable mixtures thereof. The proper fluidity can bemaintained, for example, by the formation of liposomes, by themaintenance of the required particle size in the case of dispersions orby the use of surfactants. The prevention of the action ofmicroorganisms can be brought about by various antibacterial andantifungal agents, for example, parabens, chlorobutanol, phenol, sorbicacid, thimerosal, and the like. In many cases, it will be preferable toinclude isotonic agents, for example, sugars, buffers or sodiumchloride. Prolonged absorption of the injectable compositions can bebrought about by the use in the compositions of agents delayingabsorption, for example, aluminum monostearate and gelatin.

[0090] Sterile injectable solutions are prepared by incorporating theactive compound in the required amount in the appropriate solvent withvarious of the other ingredients enumerated above, as required, followedby filter sterilization. In the case of sterile powders for thepreparation of sterile injectable solutions, the preferred methods ofpreparation are vacuum drying and the freeze drying techniques, whichyield a powder of the active ingredient plus any additional desiredingredient present in the previously sterile-filtered solutions.

[0091] For topical administration, the present compounds may be appliedin pure form, i.e., when they are liquids. However, it will generally bedesirable to administer them to the skin as compositions orformulations, in combination with a dermatologically acceptable carrier,which may be a solid or a liquid.

[0092] Useful solid carriers include finely divided solids such as talc,clay, microcrystalline cellulose, silica, alumina and the like. Usefulliquid carriers include water, alcohols or glycols orwater-alcohol/glycol blends, in which the present compounds can bedissolved or dispersed at effective levels, optionally with the aid ofnon-toxic surfactants. Adjuvants such as fragrances and additionalantimicrobial agents can be added to optimize the properties for a givenuse. The resultant liquid compositions can be applied from absorbentpads, used to impregnate bandages and other dressings, or sprayed ontothe affected area using pump-type or aerosol sprayers.

[0093] Thickeners such as synthetic polymers, fatty acids, fatty acidsalts and esters, fatty alcohols, modified celluloses or modifiedmineral materials can also be employed with liquid carriers to formspreadable pastes, gels, ointments, soaps, and the like, for applicationdirectly to the skin of the user.

[0094] Examples of useful dermatological compositions which can be usedto deliver the compounds of formula I to the skin are known to the art;for example, see Jacquet et al. (U.S. Pat. No. 4,608,392), Geria (U.S.Pat. No. 4,992,478), Smith et al. (U.S. Pat. No. 4,559,157) and Wortzman(U.S. Pat. No. 4,820,508).

[0095] Useful dosages of the compounds of formula I can be determined bycomparing their in vitro activity, and in vivo activity in animalmodels. Methods for the extrapolation of effective dosages in mice, andother animals, to humans are known to the art; for example, see U.S.Pat. No. 4,938,949.

[0096] Generally, the concentration of the compound(s) of formula I in aliquid composition, such as a lotion, will be from about 0.1-25 wt-%,preferably from about 0.5-10 wt-%. The concentration in a semi-solid orsolid composition such as a gel or a powder will be about 0.1-5 wt-%,preferably about 0.5-2.5 wt-%.

[0097] The amount of the compound, or an active salt or derivativethereof, required for use in treatment will vary not only with theparticular salt selected but also with the route of administration, thenature of the condition being treated and the age and condition of thepatient and will be ultimately at the discretion of the attendantphysician or clinician.

[0098] In general, however, a suitable dose will be in the range of fromabout 0.5 to about 100 mg/kg, e.g., from about 10 to about 75 mg/kg ofbody weight per day, such as 3 to about 50 mg per kilogram body weightof the recipient per day, preferably in the range of 6 to 90 mg/kg/day,most preferably in the range of 15 to 60 mg/kg/day.

[0099] The compound is conveniently administered in unit dosage form;for example, containing 5 to 1000 mg, conveniently 10 to 750 mg, mostconveniently, 50 to 500 mg of active ingredient per unit dosage form.

[0100] The desired dose may conveniently be presented in a single doseor as divided doses administered at appropriate intervals, for example,as two, three, four or more sub-doses per day. The sub-dose itself maybe further divided, e.g., into a number of discrete loosely spacedadministrations; such as multiple inhalations from an insufflator or byapplication of a plurality of drops into the eye.

[0101] Determination of Releasing Group Activity

[0102] To determine whether representative compounds of formula Ideliver the alkylating moiety (V) following quinone reduction,representative compounds of the invention were chemically reduced andthe reaction was monitored by ³¹P NMR. The quinone was dissolved inCH₃CN (0.3 mL, or THF for 16a) and the activating agent (glutathione, orsodium dithionite) was dissolved in cacodylate buffer (0.4 mL, 0.4 M, pH7.7). The buffer solution was added to the organic solution and the pHof the mixture was adjusted to −7.4. The reaction mixture wastransferred to a 5-mm NMR tube, and the data acquisition was started(pulse delay 30 μs). Spectra were taken every 2.5 minutes for 0.5 h,then every 5 min for 0.5 h, then every 10 min for 1 h, and time pointsfor each spectrum were assigned from the initiation of the reaction.Chemical shifts are reported relative to the TPPO reference. Thetemperature of the probe was maintained at 37° C., if necessary, usingthe Bruker variable temperature unit. The relative concentrations of theintermediates were determined by measuring the peak areas.

[0103] Naphthoquinone 15a was reduced with sodium dithionite (3 equiv,3:4 CH₃CN:0.4 M cacodylate buffer, pH˜7.4, 37° C.), and the reaction wasfollowed by ³¹P NMR. The resonance for quinone 3c at −5.3 ppmdisappeared within 5 minutes and was replaced by the resonance for thecorresponding phosphoramide mustard at −12.2 ppm. Similar results wereobtained for compounds 16a, 17, and 40a.

[0104] Other Cellular Mechanisms of Quinone Activation

[0105] The Michael addition of sulfur nucleophiles to naphthoquinones iswell known; and addition to the 3-position of the quinone might provideanother pathway for phosphorodiamidate anion release. Two possibleproducts could be predicted from the Michael addition of a nucleophileto the naphthoquinone. Addition at the 2-position leads to reversibleformation of the kinetic product; and addition at the 3-positionprovides an intermediate that could expel the nucleophile in areversible reaction, or could expel the phosphorodiamidate anion in anirreversible step. Experiments were carried out using sodiumdimethyldithiocarbamate (DDTC), which contains a highly nucleophilicsulfur that is anionic at physiologic pH. Activation of 16a with DDTC (3equiv, 3:4 THF:0.4 M cacodylate buffer, pH˜7.3, RT) was monitored using³¹P NMR. Three equivalents of nucleophile were used, assuming that oneequivalent would be consumed by the Michael addition and the other twowould be consumed by reaction with the resulting phosphorodiamidateanion. The resonance for quinone 16a (−5.7 ppm) disappeared and wasreplaced by the resonance for the phosphorodiamidate anion at −13.1 ppmwithin 5 min after addition of DDTC, confirming that nucleophilicactivation of the naphthoquinone is rapid and complete.

[0106] The experiment above demonstrate that compound 16a is activatedby a potent sulfur nucleophile. Thus, the following study was conductedto determine whether glutathione would activate compounds of formula Iin a similar way. Glutathione, the primary non-protein intracellularthiol, is responsible for maintaining the cellular redox environment andremoving potential electrophilic cytotoxins. It is overproduced (1-10 mMintracellular concentrations) in many cancer cell lines, particularlythose that have acquired resistance to anticancer drugs. Thus, thereaction of 15a with glutathione (3 equiv, 1:1.3 CH₃CN:0.4 M cacodylatebuffer, pH˜7.5, RT) was monitored by ³¹P NMR. The reaction withglutathione was essentially identical to that of DDTC; the resonance forquinone 15a had disappeared and was replaced by the resonance forphosphorodiamidate anion within 4 minutes of glutathione addition. Theresult was essentially identical when only one equivalent of glutathionewas used, again suggesting that activation of this compound is completebefore cyclization of the phosphorodiamidate anion can occur.

[0107] The cytotoxic properties of a compound of the invention can bedetermined using in vitro pharmacological models which are well known tothe art, or can be determined using Test A described below.

[0108] Test A: In Vitro Cytotoxicity (Clonogenic Survival of HT-29 andBE Cells):

[0109] The human colon carcinoma cell line HT-29 was obtained from theAmerican Type Culture Collection (ATCC), and the BE cell line wasprovided by Dr. D. Ross, University of Colorado. A modification of theprocedure described by Miribelli et al Cancer Research, 1985, 45, 32-39was used to determine the clonogenic survival of the cells. HT-29 and BEcells in exponential growth were suspended in unsupplemented Eagles MEMmedium (10 mL) at a final density of 1.7-2×10⁵ cells/mL. Unsupplementedmedium contains Minimum Essential Medium (Gibco) and HEPES (0.02 M). Thedrug stock solutions (0.1-40 mM) were prepared using either ethanol ordimethyl sulfoxide as solvent. The maximum amount of DMSO or ethanolused in the drug treatments was 1% of the total volume. Appropriatevolumes (6.5 to 100 μL) of the drug stock solution were added to fivevials of the cell suspensions, to give five different final drugconcentrations, and 100 μL of solvent was added to a sixth vial for acontrol. The treated cells were incubated for 2 h (37° C., 5% CO₂). Thecells were spun down and rinsed three times with supplemented medium (3mL) and then diluted in 5 mL of supplemented medium. Supplemented mediumis prepared by adding Fetal Bovine Serum (10%), gentamicin (0.05 mg/mL),L-glutamine (0.03 mg/mL), and sodium pyruvate (0.1 mM) to unsupplementedmedium. The cells were counted using a Coulter Counter and then platedat 2-3 different densities for each drug concentration and incubated for10 days. The colonies were stained with 0.5% crystal violet in 95%ethanol and those colonies comprised of 50 or more cells were countedusing a microscope and pen-style counter. The LC₉₉ of each compound (theconcentration at which there is a 1% cell survival) was determined byplotting the log surviving fraction vs. drug concentration. Data forrepresentative compounds of the invention is provided in Table 1. TABLE1 HT-29 BE Compound LC₉₉ (μM) LC₉₉ (μM) 15a 11 13 15b 2.1 1.0 15c 4.25.9 15d 4.1 3.3 16a 7.8 4.6 16b 1.7 1.2 17  14 7.8 40a 4.5 1.2 40b 1.50.44 40c 4.5 2.8 47  0.07 0.14

[0110] The above data suggests that compounds of formula I wherein R_(a)and/or R_(c) are alkyl are unexpectedly more cytotoxic than thecorresponding compounds wherein R_(a) and R_(c) are hydrogen. Thus,prefered compounds of the invention include compounds of formula Iwherein R_(a) and/or R_(c) are alkyl.

[0111] The growth inhibitory properties of representative compounds ofthe invention were also determined by evaluating their growth inhibitoryactivity against a series of human tumor cell lines using a 72-h drugexposure. Cell counts were measured using the MTT assay. The results aresummarized in Table 2. TABLE 2 Growth inhibitory activity of compounds1-6 IC₅₀, nM^(a) Cpd IGROV MDA231 PC-3 HT-29 PaCa-2 A498 A549 UMUC3 40a2048 3228 2559 5925 2303 4459 11181 9953 16a 18932 14656 9193 15266 441715599 4583 2823 47 494 40 66 65 25 344 23 2  4 194 649 241 215 998 39810  5 7588 210 420 93 13 2650 260 14  6a 1700 3500 170 340 6 862 203 22 6b 1473 1628 195 61 35 199 9 1

[0112] Several interesting conclusions are apparent from these data.Compounds 40a and 16a are moderately active (IC₅₀ in the low micromolarrange) against all cell lines and show relatively little selectivity fortumor cells of different tissue types. Second, compounds 3, 5, 6a and 6bshow significant selectivity and exceptional potency against PaCa-2 andUMUC3 cell lines, with IC₅₀ values in the low nanomolar range. Theseresults are especially significant because pancreatic cancer and bladdercancer are difficult tumors to treat clinically. These tumors are poorlyresponsive to most existing anticancer drugs, and therapeutic optionsare very limited.

[0113] The cytotoxic properties of a compound of the invention can alsobe determined using in vivo pharmacological models which are well knownto the art. For example, compound 16a was tested in a xenograft model.This assay consisted of injecting nude mice subcutaneously with humantumor cells. The tumors were allowed to grow until a measurable mass waspresent (22 days), and a single dose of 16a was administeredsubcutaneously. The tumor size was measured periodically and the growthinhibition of the tumor was determined. The A498₂LM cell line, asub-line of the A498 human kidney carcinoma, was used for the xenograftassay. The A498₂LM sub-line, which has a high DT-diaphorase level (1010nmol cytochrome c reduced/min/mg protein), was developed by implantingthe A498 cell line in mice and collecting the lung metastases thatformed from the original tumor. Compound 16a exhibited a 40% growthinhibition on Day 38 after a single dose of 25 mg/kg (Graph 2).

[0114] The invention will now be illustrated by the followingnon-limiting Examples wherein unless otherwise noted the followinggeneral procedures were followed.

[0115] General Procedures

[0116]¹H NMR spectra were measured on a 250 MHz Bruker NMR systemequipped with a multinuclear (¹H, ¹³C, ⁹F and ³¹P) 5-mm probe. The NMRdata acquisition/processing program MacNMR was used with the Tecmag dataacquisition system. ¹H Chemical shifts are reported in parts per millionfrom tetramethylsilane. ³¹P NMR spectra were obtained on the sameinstrument using broadband gated decoupling. Chemical shifts arereported in parts per million from a coaxial insert containing 5%phosphoric acid in H₂O. All variable temperature experiments wereconducted using a Bruker variable temperature unit.

[0117] Analtech precoated silica gel glass plates (250 microns) wereused to perform thin layer chromatography. The plates were visualizedusing UV and/or one of the following two stains: 3% phosphomolybdic acidin methanol followed by heating or 1% 4-(p-nitrobenzyl)pyridine inacetone followed by heating and treatment with 3% KOH in acetone (todetect for an alkylating moiety). Chromatographic purifications werecarried out by flash chromatography using silica gel grade 60 (230400mesh, 60 A). High performance liquid chromatography (HPLC) analyses wereperformed using a Beckman System Gold with a 126 Solvent Module, a 168Detector set to either 250 or 280 nm and an Econosphere C18 5-microncolumn (250 mm, Alltech Associates). The mobile phase was acetonitrile:0.1% trifluoroacetic acid in H₂O using the percentages indicated and aflow rate of 1 mL/min.

[0118] Elemental analyses were performed by the Purdue UniversityMicroanalysis Lab, West Lafayette, Ind. Mass spectral data was obtainedfrom the Purdue University Mass Spectrometry Service, West Lafayette,Ind., using fast atom bombardment (FAB) with a 3-nitrobenzyl alcoholmatrix.

[0119] A glass-calomel electrode on either a Radiometer pH meter or anOrion PerpHect LogR meter, model 330, was used for acidity measurements.Melting points were determined on a Mel-Temp II apparatus and areuncorrected. IR spectra were recorded on a Nicolet Magna IR 550spectrometer using either a thin film or Nujol suspension, as noted,between NaCl plates. All anhydrous reactions were carried out in eithera flame dried or oven dried flask under argon. Organic solutions wereconcentrated on a Buchi rotary evaporator.

[0120] Chemical reagents were purchased from Aldrich except NADH, NADPH,dicumarol and cytochrome c (Sigma) and ammonia (Matheson Gas). All cellculture reagents were purchased from Gibco Life Technologies. Purifiedhuman DT-diaphorase was supplied by Dr. S. Chen, City of Hope MedicalCenter, CA.

[0121] Tetrahydrofuran was distilled from sodium, with benzophenoneketyl as indicator, prior to use. Methylene chloride,diisopropylethylamine, triethylamine and acetonitrile were distilledfrom calcium hydride prior to use.

[0122] Butyllithium was purchased as a 2.5 M solution in hexanes,t-butyl hydroperoxide as a 5-6 M solution in decane, phosphorustrichloride as a 2 M solution in methylene chloride, lithium aluminumhydride as a 1.0 M solution in ether, sodium hydride as a 60% dispersionin mineral oil and lithium bis(trimethylsilyl)amide as a 1.0 M solutionin tetrahydrofuran.

[0123] Schemes For Examples 1-4

EXAMPLE 1 2-(1,4-Naphthoquinonyl)methyl N,N-bis(2-chloroethyl)phosphorodiamidate (15a)

[0124] Ceric ammonium nitrate (1.56 g, 2.85 mmol) in H₂O (10 mL) wasadded in portions over 15 min to a solution of 25a (480 mg, 1.14 mmol)in acetonitrile (30 mL). The solution was stirred at room temperaturefor 1 h and extracted with CHCl₃ (3×). The combined organic layers weredried (MgSO₄), filtered and evaporated. Column chromatography of thecrude product (6:94 MeOH:CHCl₃) afforded 15a (360 mg, 81%) as a yellowsolid; R_(f)=0.45 (6:94 MeOH:CHCl₃); mp=105-107° C.; ¹H NMR (CDCl₃): δ8.09 (m, 2H), 7.77 (m, 2H), 7.03 (t, 1H), 5.05 (m, 2H, JP-H=7.1 Hz),3.68 (t, 4H), 3.54 (m, 4H), 3.01 (bs, 2H); ³¹P NMR (CDCl₃): δ 16.42

[0125] IR (Nujol): 1664, 1630, 1589 cm⁻¹. Anal. Calcd. forC₁₅H₁₇Cl₂N₂O₄P: C, 46.06; H, 4.38; N, 7.16. Found: C, 45.72; H, 4.17; N,7.27.

[0126] The intermediate compound 25a was prepared as follows.

[0127] a. Methyl 1,4-dimethoxy-2-naphthoate (22)

[0128] Potassium carbonate (30.50 g, 0.22 mol) and MeI (27.5 mL, 0.44mol) were added to a solution of 1,4-dihydroxy-2-naphthoic acid (6.00 g,0.029 mol) in acetone (120 mL) under argon. The mixture was refluxed for48 h. Water (50 mL) was added and the mixture was extracted with CH₂Cl₂(5×). The combined organic layers were washed with H₂O (2×), dried(MgSO₄), filtered and evaporated. Column chromatography of the crudeproduct (15:85 EtOAc:hexanes) afforded 22 (7.05 g, 97%) as a greensolid; R_(f)=0.44 (15:85 EtOAc:hexanes); mp=48-50° C.; lit mp=52-55°C.²⁵; ¹H NMR (CDCl₃): δ 8.24 (m, 2H), 7.59 (m, 2H), 7.16 (s, 1H), 4.02(s, 3H), 4.01 (s, 3H), 4.00 (s, 3H).

[0129] b. 1,4-Dimethoxy-2-hydroxymethylnaphthalene (23a)

[0130] A solution of lithium aluminum hydride (29.2 mL, 29.2 mmol, 1 Min ether) in ether (30 mL) was heated to reflux under argon. A solutionof ester 22 (7.18 g, 29.2 mmol) in ether (30 mL) and THF (8 mL) wasadded dropwise over 35 min via an addition funnel. The milky yellowmixture was refluxed for 3 h and then cooled to 0° C. Methanol (10 mL)was added dropwise and the resulting clear yellow solution was stirredfor 1 h. Saturated NH₄Cl (20 mL) and aqueous HCl (5 mL, 10%) were addedand the mixture was extracted with ether (6×). The combined organiclayers were washed with saturated Na₂CO₃ (2×) and H₂O (2×), dried(MgSO₄), filtered and evaporated. Column chromatography of the crudeproduct (35:65 EtOAc:hexanes) afforded 23a (5.82 g, 92%) as a pinksolid; R_(f)=0.48 (35:65 EtOAc:hexanes); mp=69-70° C.; lit mp=68-70°C.²⁵; ¹H NMR (CDCl₃): δ 8.23 (dd, 1H), 8.03 (dd, 1H), 7.52 (m, 2H), 6.82(s, 1H), 4.89 (s, 2H), 3.99 (s, 3H), 3.92 (s, 3H), 1.95 (bs, 1H).

[0131] c. 2-(1,4-Dimethoxynaphthyl)methyl N,N-bis(2-chloroethyl)phosphorodiamidate (25a). Lithium bis(trimethylsilyl)amide (2.50 mL,2.50 mmol, 1.0 M in THF) was added dropwise via syringe to a solution ofalcohol 23a (500 mg, 2.29 mmol) in THF (10 mL) at −78° C. under argon.The resulting solution was stirred for 5 min and added dropwise viasyringe to a solution of bis(2-chloroethyl)phosphoramidic dichloride(20) (710 mg, 2.75 mmol) in THF (20 mL) at −78° C. The reaction mixturewas stirred at −78° C. for 1.5 h and then was warmed to −20° C. Gaseousammonia was passed through the reaction mixture for 10 min. The mixturewas stirred for an additional 10 min, aqueous HCl (2%, 30 mL) was addedand the mixture was extracted with EtOAc (4×). The combined organiclayers were washed with saturated NaCl (2×), dried (MgSO₄), filtered andevaporated. Column chromatography of the crude product (2:98 MeOH:EtOAc)afforded 25a (480 mg, 50%) as a yellow oil; R_(f)=0.59 (2:98MeOH:EtOAc); ¹H NMR (CDCl₃): δ 8.25 (dd, 1H), 8.06 (dd, 1H), 7.54 (m,2H), 6.85 (s, 1H), 5.33 (m, 2H, JP-H=7.8 Hz), 4.01 (s, 3H), 3.94 (s,3H), 3.65 (t, 4H), 3.48 (m, 4H), 2.77 (bs, 2H); ³¹P NMR (CDCl₃): δ15.72.

EXAMPLE 2 2-(3-Methyl-1,4-naphthoquinonyl)methyl N,N-bis(2-chloroethyl)phosphorodiamidate (15b)

[0132] Compound 15b was prepared from 25b (440 mg, 1.01 mmol) asdescribed above for 15a to give 289 mg (71%) of the product as a yellowsolid after column chromatography (3:97 MeOH:CHCl₃); R_(f)=0.16 (3:97MeOH:CHCl₃); mp=129-130° C.; ¹H NMR (CDCl₃): δ 8.11 (m, 2H), 7.76 (m,2H), 5.06 (m, 2H, JP-H=7.3 Hz), 3.66 (t, 4H), 3.47 (dt, 4H), 3.02 (bs,2H), 2.34 (s, 3H); ³¹P NMR (CDCl₃): δ 14.27; IR (Nujol): 1665, 1660,1628, 1594 cm⁻¹. Anal. Calcd. for C₁₆H₁₉Cl₂N₂O₄P: C, 47.43; H, 4.73; N,6.91. Found C, 47.59; H, 4.53; N, 6.84.

[0133] The intermediate compound 25b was prepared as follows.

[0134] a. 1,4-Dimethoxy-3-methyl-2-hydroxymethylnaphthalene (23b)

[0135] n-Butyllithium (7.3 mL, 18.3 mmol, 2.5 M in hexanes) was addeddropwise via syringe to a solution of 23a (1.00 g, 4.58 mmol) in THF (30mL) at −78° C. under argon. The solution was slowly warmed to roomtemperature and stirred for 1 h. Methyl iodide (0.34 mL, 5.50 mmol) wasadded dropwise and the solution was stirred for 20 min. Water (8 mL) wasadded and the mixture was extracted with EtOAc (4×). The combinedorganic layers were dried (MgSO₄), filtered and evaporated. Columnchromatography of the crude product (45:55 EtOAc:hexanes) afforded 23b(0.61 g, 57%) as a yellow solid; R_(f)=0.59 (45:55 EtOAc:hexanes);mp=116-117° C.; lit mp=118-121,C²⁵; ¹H NMR (CDCl₃): δ 8.07 (dd, 2H),7.51 (m, 2H), 4.94 (s, 2H), 3.98 (s, 3H), 3.88 (s, 3H), 2.52 (s, 3H),1.73 (bs, 1H).

[0136] b. 2-(3-Methyl-1,4-dimethoxynaphthyl)methylN,N-bis(2-chloroethyl)phosphorodiamidate (25b). Compound 25b wasprepared from 23b (500 mg, 2.15 mmol) as described above for 25a to give440 mg (47%) of the product as a light yellow foam after columnchromatography (1:99 MeOH:EtOAc); R_(f)=0.44 (1:99 MeOH:EtOAc); ¹H NMR(CDCl₃): δ 8.08 (m, 2H), 7.53 (m, 2H), 5.31 (m, 2H, JP-H=6.8 Hz), 3.98(s, 3H), 3.89 (s, 3H), 3.64 (t, 4H), 3.45 (dt, 4H), 2.77 (bs, 2H), 2.52(s, 3H); ³¹P NMR (CDCl₃): δ 15.65.

EXAMPLE 3 2-(3-Thiomethyl-1,4-naphthoquinonyl)methylN,N-bis(2-chloroethyl)phosphorodiamidate (15c)

[0137] Compound 15c was prepared from 25c (134 mg, 0.287 mmol) asdescribed above for 15a to give 72.0 mg (57%) of the product as aviscous orange oil after column chromatography (2:98 MeOH:CHCl₃);R_(f)=0.29 (2:98 MeOH:CHCl₃)

[0138]¹H NMR (CDCl₃): δ 8.10 (m, 2H), 7.75 (m, 2H), 5.21 (m, 2H,JP-H=6.1 Hz), 3.66 (t, 4H), 3.46 (dt, 4H), 2.97 (bs, 2H), 2.74 (s, 3H);³¹P NMR (CDCl₃): δ 15.25

[0139] IR (Nujol): 1666, 1643, 1588, 1542 cm⁻¹. Anal. Calcd. forC₁₆H₁₉Cl₂N₂O₄PS: C, 43.95; H. 4.38; N, 6.41. Found: C, 43.93; H, 4.21;N, 6.16.

[0140] The intermediate compound 25c was prepared as follows.

[0141] a. 1,4-Dimethoxy-3-thiomethyl-2-hydroxymethyl-naphthalene (23c)

[0142] n-Butyllithium (0.73 mL, 1.83 mmol, 2.5 M in hexanes) was addeddropwise to a solution of 23a (100 mg, 0.458 mmol) in THF (6 mL) at −78°C. under argon. The solution was slowly warmed to room temperature andstirred for 1 h. Dimethyl disulfide (0.050 mL, 0.55 mmol) was addeddropwise. The clear, yellow solution became cloudy during the 30 min thereaction mixture was stirred. Water (1 mL) was added and the mixture wasextracted with EtOAc (3×). The combined organic layers were washed withsaturated NaCl, dried (MgSO₄), filtered and evaporated. Columnchromatography of the crude product (20:80 EtOAc:hexanes) afforded 23c(71.2 mg, 59%, 82% based on recovered 23a, 28.0 mg) as a light greenoil; R_(f)=0.26 (20:80 EtOAc:hexanes); ¹H NMR (CDCl₃): δ 8.10 (m, 2H),7.55 (m, 2H), 5.07 (s, 2H), 4.03 (s, 3H), 3.99 (s, 3H), 2.53 (s, 3H),1.60 (bs, 1H).

[0143] b. 2-(3-Thiomethyl-1,4-dimethoxynaphthyl)methylN,N-bis(2-chloro-ethyl) phosphorodiamidate (25c). Compound 25c wasprepared from 23c (71.0 mg, 0.27 mmol) as described above for 25a togive 76.9 mg (61%) of the product as a yellow oil after columnchromatography (EtOAc); R_(f)=0.22 (EtOAc); ¹H NMR (CDCl₃): δ 8.12 (m,2H), 7.57 (m, 2H), 5.48 (d, 2H, J=4.9 Hz), 4.04 (s, 3H), 4.00 (s, 3H),3.64 (t, 4H), 3.49 (dt, 4H), 2.73 (bs, 2H), 2.51 (s, 3H); ³¹P NMR(CDCl₃): δ 14.62.

EXAMPLE 4 2-(3-Bromo-1,4-naphthoquinonyl)methylN,N-bis(2-chloroethyl)phosphorodiamidate (15d)

[0144]

[0145] Silver (II) oxide (300 mg, 2.4 mmol) was added in one portion toa solution of 25d (200 mg, 0.40 mmol) in THF (15 mL) at roomtemperature. Nitric acid (6 N, 2 mL) was added to this suspension. Themixture was stirred for 20 min at room temperature, H₂O (5 mL) was addedand the mixture was extracted with CHCl₃ (3×). The combined organiclayers were washed with saturated NaCl and NaHCO₃ (1 M), dried (MgSO₄),filtered and evaporated. Column chromatography of the crude product(2:98 MeOH:CHCl₃ until less polar impurities are removed, then 6:94MeOH:CHCl₃) afforded 15d (156 mg, 83%) as a yellow solid; R_(f)=0.36(6:94 MeOH:CHCl₃); mp=134-135° C.; ¹H NMR (CDCl₃): δ 8.18 (m, 2H), 7.80(m, 2H), 5.21 (m, 2H, JP-H=6.5 Hz), 3.66 (t, 4H), 3.48 (dt, 4H), 2.97(bs, 2H).

[0146]³¹P NMR (CDCl₃): δ 15.32; IR (Nujol): 1680, 1675, 1659, 1605, 1589cm⁻¹. Anal. Calcd. for C₁₅H₁₆BrCl₂N₂O₄P: C, 38.33; H, 3.43; N, 5.96.Found: C, 38.68; H, 3.42; N, 5.83.

[0147] The intermediate compound 25d was prepared as follows.

[0148] a. 1,4-Dimethoxy-3-bromo-2-hydroxyrmethylnaphthalene (23d)

[0149] n-Butyllithium (2.9 mL, 7.33 mmol, 2.5 M in hexanes) was addeddropwise to a solution of 23a (400 mg, 1.83 mmol) in THF (25 mL) at −78°C. under argon. The reaction mixture was slowly warmed to roomtemperature and stirred for 1 h. 1,2-dibromotetrafluoroethane (0.26 mL,2.20 mmol) was added dropwise and the solution was stirred for 1 h.Water (10 mL) was added and the mixture was extracted with EtOAc (3×).The combined organic layers were dried (MgSO₄), filtered and evaporated.Column chromatography of the crude product (35:65 EtOAc:hexanes)afforded 23d (273 mg, 50%, 63% based on recovered 23a, 84 mg) as ayellow solid; R_(f)=0.62 (35:65 EtOAc:hexanes); mp=115-117° C.; ¹H NMR(CDCl₃): δ 8.11 (m, 2H), 7.57 (m, 2H), 5.03 (s, 2H), 4.02 (s, 3H), 3.99(s, 3H), 2.36 (bs, 1H).

[0150] b. 2-(3-Bromo-1,4-dimethoxynaphthyl)methylN,N-bis(2-chloroethyl)-phosphorodiamidate (25d). Compound 25d wasprepared from 23d (270 mg, 0.91 mmol) as described above for 25a to give353 mg (78%) of the product as a white solid after column chromatography(EtOAc); R_(f)=0.17 (EtOAc); mp=112-1140C; ¹H NMR (CDCl₃): δ 8.12 (m,2H), 7.60 (m, 2H), 5.40 (m, 2H, JP-H=5.9 Hz), 4.02 (s, 3H), 4.00 (s,3H), 3.66 (t, 4H), 3.48 (dt, 4H), 2.82 (bs, 2H); ³¹P NMR (CDCl₃): δ14.38.

Scheme for Examples 5 and 6

[0151]

EXAMPLE 5 2-(1,4-Naphthoquinonyl)methyl N,N-bis(2-bromoethyl)phosphorodiamidate (16a)

[0152] Ceric ammonium nitrate (1.83 g, 3.33 mmol) in H₂O (15 mL) wasadded in portions over 15 min to a solution of 26a (680 mg, 1.33 mmol)in CH₃CN (60 mL). The reaction was stirred at room temperature for 1 hand extracted with CHCl₃ (3×). The combined organic layers were dried(MgSO₄), filtered and evaporated. Column chromatography of the crudeproduct (2:98 MeOH:EtOAc) afforded 16a (550 mg, 86%) as a yellow solid;R_(f)=0.51 (2:98 MeOH:EtOAc); mp=118-119° C.; ¹H NMR (CDCl₃): δ 8.10 (m,2H), 7.77 (m, 2H), 7.03 (t, 1H), 5.05 (m, 2H, JP-H=7.0 Hz), 3.55 (m,8H), 2.98 (bs, 2H); ³¹P NMR (CDCl₃): δ 16.18; IR (Nujol): 1664, 1627,1591 cm⁻¹. Anal. Calcd. for C₁₅H₁₇Br₂N₂O₄P: C, 37.53; H, 3.57; N, 5.83.Found: C, 37.80; H, 3.57; N, 5.56.

[0153] The intermediate compound 19 was prepared as follows.

[0154] a. bis(2-Bromoethyl)amine hydrobromide (18Br)

[0155] Hydrobromic acid (75 mL, 0.45 mol, 48% by wt) was added slowly,with stirring, to diethanolamine (11.0 g, 0.10 mol) at 0° C. Thereaction mixture was heated to reflux and then distilled through a 14/20vigreaux column. The distillate temperature was 100° C. for the first 15mL collected and 125° C. for the remainder. After a total of 40 mL ofdistillate was collected, additional 48% HBr (50 mL, 0.30 mol) was addedand 50 mL of distillate was collected. Hydrobromic acid (25 mL, 48%) wasadded and the reaction mixture was refluxed overnight. The reactionmixture was then distilled (−30 mL collected) and the still pot residuewas poured into acetone at −78° C. The white solid that precipitated(10.3 g, 22% diethanolamine hydrobromide and 78% product 18Br asdetermined by ¹H NMR) was collected by filtration. This impure productcan either be used directly in the subsequent phosphorylation reactionor can be purified by repeated recrystallization from acetone. Thisrecrystallization procedure actually crystallizes out diethanolaminehydrobromide, so that purer product 18Br is recovered from the filtrateupon successive recrystallization attempts; ¹H NMR of 18Br (D₂O): δ 3.55(t, 4H), 3.45 (t, 4H).

[0156]¹H NMR of diethanolamine hydrobromide (D₂O): δ 3.78 (t, 4H), 3.12(t, 4H).

[0157] b. bis(2-Bromoethyl)phosphoramidic dichloride (19):

[0158] Phosphorus oxychloride (0.30 mL, 3.2 mmol) was added slowly to asuspension of hydrobromide salt 18Br (1.00 g, 3.2 mmol) in CH₂Cl₂ (22mL) at −40° C. under argon. Triethylamine (1.20 mL, 8.6 mmol) was addeddropwise via syringe over 5 min while the reaction mixture wasvigorously stirred to avoid local heating. The cloudy white reactionmixture was warmed to 0° C. over 2.5 h and stirred at 0° C. for 4 h.Saturated ammonium chloride (8 mL) was added and the mixture wasextracted with CH₂Cl₂ (3×). The combined organic layers were dried(MgSO₄), filtered and evaporated. Column chromatography of the crudeproduct (20:80 EtOAc:hexanes) afforded 19 (0.76 g, 68%) as a whitesolid; R_(f)=0.56 (20:80 EtOAc:hexanes); mp=43-440C; lit mp=39-40° C.²⁵;¹H NMR (CDCl₃): δ 3.73 (dt, 4H), 3.55 (t, 4H); ³¹P NMR (CDCl₃): δ 16.48.

[0159] The intermediate compound 26a was prepared as follows.

[0160] c. 2-(1,4-Dimethoxynaphthyl)methyl N,N-bis(2-bromoethyl)phosphorodiamidate (26a). Lithium bis(trimethylsilyl)amide (6.05 mL,6.05 mmol, 1 M in THF) was added dropwise via syringe to a solution ofalcohol 23a (1.20 g, 5.50 mmol) in THF (20 mL) at −780C under argon. Theresulting solution was stirred for 5 min and then added dropwise to asolution of bis(2-bromoethyl)-phosphoramidic dichloride (19) (2.29 g,6.60 mmol) in THF (50 mL) at −78° C. The reaction mixture was stirred at−78° C. for 1.5 h and then warmed to −20° C. Gaseous ammonia was passedthrough the reaction mixture for 10 min. The mixture was stirred for 10min, aqueous HCl (2%, 70 mL) was added and the mixture was extractedwith EtOAc (3×). The combined organic layers were washed with saturatedNaCl (2×), dried (MgSO₄), filtered and evaporated. Column chromatographyof the crude product (70:30 EtOAc:hexanes) afforded 26a (1.20 g, 43%) asa white solid; R_(f)=0.33 (70:30 EtOAc:hexanes); mp=112-113° C.; ¹H NMR(CDCl₃): δ 8.23 (dd, 1H), 8.06 (dd, 1H), 7.54 (m, 2H), 6.84 (s, 1H),5.26 (m, 2H, JP-H=8.1 Hz), 4.01 (s, 3H), 3.94 (s, 3H), 3.50 (m, 8H),2.78 (bs, 2H); ³¹P NMR (CDCl₃): δ 15.67.

EXAMPLE 6 2-(3-Methyl-1,4-naphthoquinonyl)methyl N,N-bis(2-bromoethyl)phosphorodiamidate (16b)

[0161] Compound 16b was prepared from 23b (470 mg, 0.897 mmol) asdescribed above for 16a to give 230 mg (52%) of the product as a yellowsolid after column chromatography (6:94 MeOH:CHCl₃); R_(f)=0.28 (6:94MeOH:CHCl₃); mp=117-119° C.; ¹H NMR (CDCl₃): δ 8.12 (m, 2H), 7.76 (m,2H), 5.06 (m, 2H, JP-H=7.6 Hz), 3.50 (m, 8H), 3.02 (bs, 2H), 2.34 (s,3H); 31P NMR (CDCl₃): δ 15.72; IR (Nujol): 1662, 1626, 1594, 1568 cm⁻¹;HPLC (50:50 CH₃CN:0.1% TFA/1H₂O): 4.78 min, 95.6%; FAB MS: Calcd. forC₁₆H₁₉Br₂N₂O₄P: (M+H)+492.9527; Found 492.9542.

[0162] The intermediate compound 23b was prepared as follows.

[0163] a. 2-(3-Methyl-1,4-dimethoxynaphthyl)methylN,N-bis(2-bromoethyl)phosphorodiamidate (26b). Compound 26b was preparedfrom 23b (480 mg, 2.07 mmol) as described above for 26a to give 480 mg(44%) of the product as a viscous yellow oil after column chromatography(2:98 MeOH:EtOAc); R_(f)=0.63 (2:98 MeOH:EtOAc); ¹H NMR (CDCl₃): δ 8.09(d, 2H), 7.41 (m, 2H), 5.31 (m, 2H, JP-H=7.1 Hz), 3.98 (s, 3H), 3.89 (s,3H), 3.48 (m, 8H), 2.77 (bs, 2H), 2.53 (s, 3H); ³¹P NMR (CDCl₃): δ15.32.

EXAMPLE 7 2-(1,4-Naphthoquinonyl)methyl bis[N-(2-chloroethyl)]phosphorodiamidate (17)

[0164]

[0165] Ceric ammonium nitrate (0.20 g, 0.37 mmol) in H₂O (2 mL) wasadded in portions over 10 min to a solution of 28 (62.0 mg, 0.15 mmol)in CH₃CN (5 mL). The solution was stirred at room temperature for 1 h.Water (3 mL) was added and the mixture was extracted with CHCl₃ (3×).The combined organic layers were dried (MgSO₄), filtered and evaporated.Column chromatography of the crude product (78:22 CHCl₃:Acetone untilless polar impurities are removed, then 67:33 CHCl₃:Acetone) afforded 17(42.4 mg, 74%) as a yellow solid; R_(f)=0.24 (67:33 CHCl₃:Acetone);mp=109-1100C; ¹H NMR (CDCl₃): δ 8.10 (m, 2H), 7.77 (m, 2H), 7.03 (t, 1H,J=1.9 Hz), 5.04 (dd, 2H, J=1.9 and 6.8 Hz), 3.65 (t, 4H), 3.36 (dt, 4H),3.20 (bs, 2H); ³¹P NMR (CDCl₃): δ 14.83; IR (Nujol): 1662, 1633, 1594cm⁻¹. Anal. Calcd. for C₁₅H₁₇Cl₂N₂O₄P: C, 46.06; H, 4.38; N, 7.16.Found: C, 46.04; H, 4.08; N, 6.78.

[0166] The intermediate compound 28 was prepared as follows.

[0167] a. 2-(1,4-Dimethoxynaphthyl)methyl bis[N-(2-chloroethyl)]phosphorodiamidate (28):

[0168] Phosphorus oxychloride (0.043 mL, 0.46 mmol) was added to asuspension of N-(2-chloroethyl)amine hydrochloride (112 mg, 0.96 mmol)in CH₂Cl₂ (10 mL) at −78° C. under argon. Diisopropylethylamine (0.34mL, 1.92 mmol) was added dropwise and the reaction mixture was warmed toroom temperature over 2 h and stirred for 7 h. After the phosphorylationreaction was completed, n-butyllithium (0.18 mL, 0.46 mmol, 2.5 M inhexanes) was added to a solution of alcohol 23a (100 mg, 0.46 mmol) inCH₂Cl₂ (5 mL) in a second flask at −78° C. under argon. The resultingsolution was stirred for 15 min. The contents of the flask containingthe phosphorylating reagent were added via syringe to the secondsolution with vigorous stirring at −78° C. The reaction mixture wasslowly warmed to room temperature, stirred overnight, and thenconcentrated under reduced pressure until the volume was reduced to athird. The salts were separated and the crude product was purified bycolumn chromatography (78:22 CHCl₃:Acetone until less polar impuritiesare removed, then 67:33 CHCl₃:Acetone) to afford 28 (62.0 mg, 32%, 87%based on recovered 23a, 63.0 mg) as a light yellow oil; R_(f)=0.31(78:22 CHCl₃:Acetone); ¹H NMR (CDCl₃): δ 8.24 (dd, 1H), 8.06 (dd, 1H),7.55 (m, 2H), 6.84 (s, 1H), 5.25 (d, 2H, J=7.9 Hz), 4.01 (s, 3H), 3.94(s, 3H), 3.59 (t, 4H), 3.28 (dt, 4H), 3.04 (bs, 2H); ³¹P NMR (CDCl₃): δ14.72.

Scheme For Examples 8-10

[0169]

EXAMPLE 8 2-(1,4-Naphthoquinonyl)methylbis[N-methyl-N-(2-bromoethyl)]phosphorodiamidate (40a)

[0170] Ceric ammonium nitrate (510 mg, 0.93 mmol) in H₂O (5 mL) wasadded in portions over 10 min to a solution of 41a (200 mg, 0.372 mmol)in CH₃CN (17 mL). The solution was stirred at room temperature for 1 hand extracted with CHCl₃ (3×). The combined organic layers were washedwith H₂O and saturated NaCl, dried (MgSO₄), filtered and evaporated.Column chromatography of the crude product (2:98 MeOH:CH₂Cl₂) afforded40a (164 mg, 87%) as a viscous brown oil; R_(f)=0.16 (2:98 MeOH:CH₂Cl₂);¹H NMR (CDCl₃): δ 8.10 (m, 2H), 7.77 (m, 2H), 7.03 (t, 1H, J=2.0 Hz),5.04 (dd, 2H, J=2.0 and 6.4 Hz), 3.49 (m, 8H), 2.78 (d, 6H, J=9.7 Hz);³¹P NMR (CDCl₃): δ 18.00; IR (neat): 1664, 1632, 1595 cm⁻¹.

[0171] Anal. Calcd. for C₁₇H₂₁Br₂N₂O₄P: C, 40.18; H, 4.17; N, 5.51.Found: C, 40.36; H, 4.09; N, 5.18.

[0172] The intermediate compound 41a was prepared as follows.

[0173] a. 2-(1,4-Dimethoxynaphthyl)methyl bis[N-methyl-N-(2-bromoethyl)]phosphorodiamidate (41a). A solution of alcohol 23a (100 mg, 0.46 mmol)in CH₂Cl₂ (2 mL) and CH₃CN (2 mL) was cooled to −70° C. and stirredunder argon. Phosphorus trichloride (0.23 mL, 0.46 mmol, 2 M in CH₂Cl₂)was added dropwise followed by the dropwise addition ofdiisopropylethylamine (0.08 mL, 0.46 mmol). The reaction mixture wasstirred for 25 min at −70° C. A solution ofN-methyl-N-(2-bromoethyl)amine hydrobromide (200 mg, 0.92 mmol) in CH₃CN(2.5 mL) was added slowly via syringe, followed by additionaldiisopropylethylamine (0.32 mL, 1.84 mmol). The temperature was raisedto −60 to −50° C. and the reaction was stirred for 1.25 h. t-Butylhydroperoxide (1.1 mL, 5-6 M in decane) was added and the reactionmixture was warmed to −40 to −20° C. and stirred for 30 min. Water (3mL) was added and the mixture was warmed to room temperature andextracted with CH₂Cl₂ (4×). The combined organic layers were washed withH₂O and saturated NaCl, dried (MgSO₄), filtered and evaporated. Columnchromatography of the crude product (2:98 MeOH:EtOAc) afforded 41a (190mg, 77%) as a light yellow oil; R_(f)=0.33 (2:98 MeOH:EtOAc); ¹H NMR(CDCl₃): δ 8.24 (dd, 1H), 8.06 (dd, 1H), 7.54 (m, 2H), 6.87 (s, 1H),5.24 (d, 2H, J=7.4 Hz), 4.01 (s, 3H), 3.93 (s, 3H), 3.43 (m, 8H), 2.71(d, 6H, J=9.7 Hz); ³¹P NMR (CDCl₃): δ 17.60.

EXAMPLE 9 2-(3-Methyl-1,4-naphthoquinonyl)methylbis[N-methyl-N-(2-bromoethyl)] phosphorodiamidate (40b)

[0174] Compound 40b was prepared from 41b (130 mg, 0.235 mmol) asdescribed above for 40a to give 99 mg (81%) of the product as a yellowoil after column chromatography (3:97 MeOH:CH₂Cl₂); R_(f)=0.20 (3:97MeOH:CH₂Cl₂); ¹H NMR (CDCl₃): δ 8.12 (m, 2H), 7.75 (m, 2H), 5.04 (d, 2H,J=6.4 Hz), 3.43 (m, 8H), 2.71 (d, 6H, J=9.7 Hz), 2.35 (s, 3H); ³¹P NMR(CDCl₃): δ 17.77; IR (neat): 1663, 1626, 1595 cm⁻¹. Anal. Calcd. forC₁₈H₂₃Br₂N₂O₄P: C, 41.40; H, 4.44; N, 5.36. Found: C, 41.31; H, 4.28; N,5.13.

[0175] The intermediate compound 41b was prepared as follows.

[0176] a. 2-(3-Methyl-1,4-dimethoxynaphthyl)methylbis[N-methyl-N-(2-bromoethyl)] phosphorodiamidate (41b). Compound 41bwas prepared from alcohol 23b (100 mg, 0.431 mmol) andN-methyl-N-(2-bromoethyl)amine hydrobromide as described above for 41ato give 135 mg (57%) of the product as a yellow oil after columnchromatography (2:98 MeOH:EtOAc); R_(f)=0.37 (2:98 MeOH:EtOAc); ¹H NMR(CDCl₃): δ 8.09 (dd, 2H), 7.52 (m, 2H), 5.28 (d, 2H, J=5.8 Hz), 3.97 (s,3H), 3.89 (s, 3H), 3.41 (m, 8H), 2.67 (d, 6H, J=9.7 Hz), 2.53 (s, 3H);³¹P NMR (CDCl₃): δ 17.28.

EXAMPLE 10 2-(1,4-Naphthoquinonyl)methylbis[N-methyl-N-(2-chloroethyl)]phosphorodiamidate (40c)

[0177] Compound 40c was prepared from 41c (130 mg, 0.289 mmol) asdescribed above for 40a to give 105 mg (87%) of the product as a brownoil after column chromatography (2:98 MeOH:EtOAc); R_(f)=0.55 (2:98MeOH:EtOAc); ¹H NMR (CDCl₃): δ 8.10 (m, 2H), 7.77 (m, 2H), 7.03 (t, 1H,J=2.0), 5.03 (dd, 2H, J=2.0 and 6.4 Hz), 3.66 (t, 4H), 3.42 (m, 4H),2.78 (d, 6H, J=9.9 Hz); ³¹P NMR (CDCl₃): δ 18.21 ppm; IR (neat): 1664,1633, 1595 cm⁻¹. Anal. Calcd. for C₁₇H₂₁Cl₂N₂O₄P: C, 48.70; H, 5.05; N,6.68. Found: C, 48.36; H, 5.02; N, 6.35.

[0178] The intermediate compound 41c was prepared as follows.

[0179] a. 2-(1,4-Dimethoxynaphthyl)methylbis[N-methyl-N-(2-chloroethyl)]phosphorodiamidate (41c). Compound 41cwas prepared from alcohol 23a (100 mg, 0.458 mmol) andN-methyl-N-(2-chloroethyl)amine hydrochloride as described above for 41ato give 132 mg (64%) of the product as a light yellow oil after columnchromatography (2:98 MeOH:EtOAc); R_(f)=0.33 (2:98 MeOH:EtOAc); ¹H NMR(CDCl₃): δ 8.24 (dd, 1H), 8.06 (dd, 1H), 7.54 (m, 2H), 6.88 (s, 1H),5.24 (d, 2H, J=7.3 Hz), 4.00 (s, 3H), 3.93 (s, 3H), 3.63 (t, 4H), 3.37(dt, 4H), 2.72 (d, 6H, J=9.6 Hz); ³¹P NMR (CDCl₃): δ 17.91.

EXAMPLE 11 3-(5-Methoxy-1-methyl-4,7-indolequinonyl)-methylbis[N-methyl-N-(2-bromoethyl)] phosphorodiamidate (47)

[0180]

[0181] A solution of alcohol 42 (50 mg, 0.23 mmol) in CH₂Cl₂ (3 mL) andCH₃CN (2 mL) was cooled to −70° C. and stirred under argon. Phosphorustrichloride (0.115 mL, 0.23 mmol, 2 M in CH₂Cl₂) was added dropwisefollowed by the dropwise addition of diisopropylethylamine (0.04 mL,0.23 mmol). The solution was stirred for 15 min andN-methyl-N-(2-bromoethyl)amine hydrobromide (99 mg, 0.46 mmol) in CH₃CN(1.5 mL) was added slowly via syringe followed by the dropwise additionof diisopropylethylamine (0.16 mL, 0.92 mmol). The mixture was stirredfor 2 h below −60° C. t-Butyl hydroperoxide (0.23 mL, 5-6 M in decane)was added and the solution was warmed to −40 to −20° C. and stirred for1 hr. Water (3 mL) was added and the mixture was extracted with CH₂Cl₂(4×). The combined organic layers were washed with H₂O and saturatedNaCl, dried (MgSO₄), filtered and evaporated. Column chromatography ofthe crude product (10:90 EtOH:ether) afforded 47 (25 mg, 20%) as anorange oil; R_(f)=0.48 (10:90 EtOH:ether); ¹H NMR (CDCl₃): □ 6.91 (s,1H), 5.68 (s, 1H), 5.14 (d, 2H, J=7.5 Hz), 3.96 (s, 3H), 3.83 (s, 3H),3.42 (m, 8H), 2.72 (d, 6H, J=9.7 Hz); ³¹P NMR (CDCl₃): □ 16.51; IR(neat): 1673, 1643, 1597, 1511 cm⁻¹; HPLC (40:60 CH₃CN:0.1% TFA/H₂O):8.78 min, 95.9%; FAB MS: Calcd. for C₁₇H₂₄Br₂N₃O₅P: (M+H)+539.9900;Found: 539.9901.

[0182] The intermediate compound 42 was prepared as follows.

[0183] a. 5-Methoxy-1-methylindole-3-carboxaldehyde (43)

[0184] 5-Methoxyindole-3-carboxaldehyde (300 mg, 1.71 mmol) was added inportions over 5 min to a suspension of sodium hydride (82 mg, 2.05 mmol,60% dispersion in mineral oil) in DMF (8 mL) stirring under argon. Themixture was stirred for 30 min, methyl iodide (0.13 mL, 2.05 mmol) wasadded and the mixture was stirred for 1 h. Sodium bicarbonate (10%, 40mL) was added and the mixture was extracted with EtOAc (4×). Thecombined organic layers were washed with sodium bicarbonate (10%, 2×)and saturated NaCl, dried (MgSO₄), filtered and evaporated. Columnchromatography of the crude product (50:50 EtOAc:hexanes) afforded 43(320 mg, 99%) as a light yellow solid; R_(f)=0.35 (50:50 EtOAc:hexanes);mp=130-1320C; lit mp=132-133° C.⁵³; ¹H NMR (CDCl₃): δ 9.95 (s, 1H), 7.79(d, 1H, J=2.4 Hz), 7.62 (s, 1H), 7.25 (d, J=8.8 Hz), 6.96 (dd, 1H, J=2.4and 8.9 Hz), 3.90 (s, 3H), 3.85 (s, 3H).

[0185] b. 5-Methoxy-1-methyl-4-nitroindole-3-carboxaldehyde (44)

[0186] A mixture of concentrated HNO₃ (2 mL) in AcOH (11 mL) was addeddropwise over 40 min to a solution of 43 (1.08 g, 5.71 mmol) in AcOH (70mL) at 0° C. The mixture was warmed to room temperature, stirredovernight, and poured over ice. The product (44) was collected byfiltration, washed with H₂O, and dried to give 1.18 g (88%) of a yellowsolid; R_(f)=0.49 (EtOAc); mp=195-197° C.; lit mp=197-198° C.⁵³; ¹H NMR(CDCl₃): δ 9.84 (s, 1H), 7.83 (s, 1H), 7.45 (d, 1H, J=9.0 Hz), 7.11 (d,1H, J=9.1 Hz), 3.96 (s, 3H), 3.90 (s, 3H).

[0187] c. 4-amino-5-methoxy-1 methylindole-3-carboxaldehyde (45)

[0188] Tin powder (2.65 g, 22.3 mmol) was added to a suspension ofnitroindole 44 (600 mg, 2.56 mmol) in EtOH (90 mL). Hydrochloric acid (3M, 36 mL) was added and the mixture was stirred at room temperature for2 h. The reaction mixture was decanted from the excess tin and added inportions to saturated sodium bicarbonate (200 mL). The mixture wasextracted with EtOAc (4×) and the combined extracts were washed withsodium bicarbonate (1 M, 3×) and saturated NaCl (2×), dried (MgSO₄),filtered and evaporated. Compound 45 was isolated as a dark yellow oil(513 mg, 98%) and used in the following reaction without furtherpurification; R_(f)=0.67 (EtOAc); ¹H NMR (CDCl₃): δ 9.61 (s, 1H), 7.55(s, 1H), 6.94 (d, 1H, J=8.6 Hz), 6.53 (d, 1H, J=8.6 Hz), 5.79 (bs, 2H),3.88 (s, 3H), 3.76 (s, 3H).

[0189] d. 3-Formyl-5-methoxy-1-methylindole-4,7-dione (46)

[0190] Potassium nitrosodisulfonate (1.85 g, 6.89 mmol) in H₂O (19 mL)was added to a solution of amine 45 (352 mg, 1.72 mmol) in acetone (37mL). Sodium phosphate buffer (0.4 M, pH 6, 10 mL) was added and thereaction mixture was stirred at room temperature for 1.5 h. The acetonewas removed under reduced pressure and the yellow solid was collected byfiltration and washed with H₂O. The solid was taken up in warm EtOAc(100 mL) and a small amount of an insoluble solid was removed byfiltration. The filtrate was rotovaped and the product was passedthrough a plug of silica gel (100:10:0.5 CHCl₃:EtOAc:MeOH) to afford 46(310 mg, 82%) as a yellow solid; R_(f)=0.44 (100:10:0.5CHCl₃:EtOAc:MeOH); ¹H NMR (CDCl₃): δ 10.41 (s, 1H), 7.45 (s, 1H), 5.77(s, 1H), 4.03 (s, 3H), 3.87 (s, 3H).

[0191] e. 3-Hydroxymethyl-5-methoxy-1-methylindole-4,7-dione (42)

[0192] Aldehyde 46 (50 mg, 0.228 mmol) was added to anhydrous MeOH (30mL) that had been degassed by bubbling with argon for 1.5 h. Thesuspension was degassed with argon for 15 min, NaBH₄ (65 mg, 1.71 mmol)was added and the reaction mixture was stirred for 2 h. A persistentlight yellow color indicated that the solvent was thoroughly degassedand the hydroquinone had formed. The solution turned dark orangefollowing air-oxidation to the quinone. The MeOH was removed underreduced pressure and the residue was taken up in CH₂Cl₂ (30 mL), washedwith H₂O (2×) and saturated NaCl, dried (MgSO₄), filtered andevaporated. Column chromatography of the crude product (EtOAc) afforded42 (24.2 mg, 48%) as a bright orange solid; R_(f)=0.57 (EtOAc);mp=182-184° C.; lit. mp=185-186° C.⁵³; ¹H NMR (CDCl₃): δ 6.71 (s, 1H),5.69 (s, 1H), 4.64 (s, 2H), 3.94 (s, 3H), 3.85 (s, 3H).

EXAMPLE 12 3-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylN,N-bis(2-bromoethyl)phosphorodiamidate (4)

[0193]

[0194] LHMDS (0.25 mL, 1.0M in THF) was added dropwise to a solution of3-hydroxymethyl-5-methoxy-1-methylindole-4,7-dione 12 (50 mg, 0.23 mmol)in THF (10 mL). The solution was stirred for 10 min at −78° C. Asolution of Cl₂P(O)N(CH₂CH₂Br)₂ (87 mg, 0.25 mmol) in THF (5 mL) wasadded all at once to the alkoxide, and the resulting solution wasstirred for 1.5 h at −78° C. The solution was warmed to −20° C. andammonia gas was bubbled through the reaction mixture for 6 min. Themixture was stirred for an additional 7 min and then added to CH₂Cl₂/H₂Oand extracted (CH₂Cl₂, 3×). The combined organic layers were dried overNa₂SO₄ and evaporated. Column chromatography of the crude product(EtOAc/Acetone) afforded 4 (17.8 mg, 30% based on recovered startingmaterial) as a yellow solid; R_(f)=0.47 (50% EtOAc/Acetone); ¹H NMR(CDCl₃): δ 6.88 (1H, s), 5.71 (1H, s), 5.04 (2H, m), 3.96 (3H, s), 3.84(3H, s), 3.47 (8H, m), 3.22 (2H, bs); ³¹P NMR (CDCl₃): 6-9.99(Ref=TPPO); HPLC (40% CH₃CN/0.1% TFA H₂O): 6.97 min.

[0195] The intermediate compound 12 was prepared as follows.

[0196] a. 5-Methoxy-1-methylindole-2-carboxylic acid methyl ester (8):

[0197] A solution of 5-methoxyindole-2-carboxylic acid (2.00g, 10.46mmol) in anhydrous DMF (28 mL) was added to sodium hydride (1.68g, 41.84mmol, 60% dispersion in mineral oil) at 0° C., under an argonatmosphere. The mixture was stirred for 5 min, dimethyl sulfate (2.96mL, 31.38 mmol) was added, and the reaction was stirred at roomtemperature for 48 h. HCl (2M) was added and the resulting mixtureextracted with CH₂Cl₂ (3×). The combined organic layers were dried overNa₂SO₄ and evaporated. Column chromatography of the crude product (25%EtOAc/Hexanes) afforded 8 (1.77 g, 78%) as an ivory solid; R_(f)=0.84(25% EtOAc/hexanes); ¹H NMR (CDCl₃): δ 7.33 (1H, s), 7.30 (1H, d), 7.20(1H, s), 7.14 (1H, d), 4.05 (3H, s), 3.94 (3H, s), 3.90 (3H, s).

[0198] b. 5-Methoxy-1-methyl-4-nitroindole-2-carboxylic acid methylester (9)

[0199] A solution of concentrated HNO₃ (2 mL) in AcOH (9 mL) was addedto a solution of 8 (830 mg, 3.79 mmol) in AcOH (54 mL) at 0° C.Following the addition, the reaction mixture was warmed to roomtemperature and stirred for 2 h. The reaction mixture was poured overice, filtered and the precipitate washed with H₂O. The precipitate wasdissolved in CH₂Cl₂ and filtered through a short column of silica gel toafford 9 (780 mg, 78%) as a yellow solid; R_(f)=0.53 (50%EtOAc/hexanes); ¹H NMR (CDCl₃): δ 7.58 (1H, d), 7.55 (1H, s), 7.18 (1H,d), 4.11 (3H, s), 4.02 (3H, s), 3.94 (3H, s).

[0200] c. 4-Amino-5-methoxy-1-methylindole-2-carboxylic acid methylester (10):

[0201] To a solution of 9 (11.0g, 3.78 mmol) in anhydrous methanol (90mL) was added 110% Pd/C (150 mg) suspended in anhydrous methanol (15 mL)followed by ammonium formate (1.10 g, 17.4 mmol). The reaction mixturewas stirred for 1 h, filtered through celite and the methanol removed.The residue was taken up in CH₂Cl₂/H₂O and extracted with CH₂Cl₂ (3×).The combined organic layers were dried over Na₂SO₄ and evaporated toafford 10 (595 mg, 67%) as a brown solid which was used without furtherpurification; ¹H NMR (CDCl₃): δ 7.21 (1H, s), 7.06 (1H, d), 6.72 (1H,d), 4.01 (3H, s), 3.90 (3H, s), 3.88 (3H, s).

[0202] d. 4-Amino-2-hydroxymethyl-5-methoxy-1-methylindole (11)

[0203] Lithium aluminum hydride (5.84 mL, 5.84 mmol, 1.0 M solution inEt₂O) was added to a solution of 10 (595 mg, 2.59 mmol) in THF (12 mL)under argon. The reaction was then heated at reflux for 15 min, quenchedby careful addition of H₂O followed by 1 M NaOH, then extracted withCH₂Cl₂ (3×). The combined organic layers were dried over Na₂SO₄ andevaporated to afford 11 (490 mg, 94%) as a brown oil which was usedwithout further purification; ¹H NMR (CDCl₃): δ 6.95 (1H, d), 6.71 (1H,d), 6.34 (1H, s), 4.77 (2H, s), 3.87 (3H, s), 3.74 (3H, s).

[0204] e. 2-Hydroxymethyl-5-methoxy-1-methylindole-4,7-dione (12)

[0205] A solution of potassium nitrosodisulfonate (KSO₃)₂NO (590 mg, 2.2mmol) in sodium phosphate buffer (0.4 M, pH=6, 13 mL) was added to asolution of 11 (130 mg, 0.63 mmol) in acetone (8 mL). The reactionmixture was stirred for 1 h at room temperature, ethyl acetate and waterwere added, and the aqueous layer was extracted (EtOAc, 3×). Thecombined organic layers were dried over Na₂SO₄ and evaporated. Columnchromatography of the crude product (EtOAc) afforded 12 (122 mg, 87%) asan orange solid; R_(f)=0.62 (EtOAc); ¹H NMR (CDCl₃): δ 6.58 (1H, s),5.67 (1H, s), 4.68 (2H, s), 4.03 (3H, s), 3.83 (3H, s).

EXAMPLE 13 2-(5-Methoxy-1-methyl-4,7-indolequinonyl)methyl bis[N-methyl-

[0206]

[0207] N-(2-bromoethyl)]phosphorodiamidate (5):

[0208] PCl₃ (0.12 ml, 2.0 M in CH₂Cl₂) was added dropwise to a solutionof 12 (50 mg, 0.23 mmol) in CH₂Cl₂ (3 mL) and CH₃CN (2 mL) at −78° C.,followed by the dropwise addition of i-Pr₂NEt (0.04 mL, 0.23 mmol). Thereaction was stirred under argon at −78° C. After 15 min,methylbromoethylamine hydrobromide (99 mg, 0.46 mmol) in CH₃CN (1.5 mL)was added, followed by the dropwise addition of i-Pr₂NEt (0.16 mL, 0.92mmol). The mixture was stirred for 2 h, t-BuOOH (0.12 mL, 5-6M indecane) was added, and the reaction mixture was warmed to −20° C. andstirred for 1 h. Water (3 mL) was added and the mixture was extractedwith CH₂Cl₂ (3×). The combined organic extracts were washed with water,dried over Na₂SO₄, and concentrated. Reverse phase column columnchromatography (45% MeOH/H₂O) afforded 5 (2.4 mg, 2%) as a yellow solid;R_(f)=0.33 (5% MeOH/EtOAc); ¹H NMR (CDCl₃): δ 6.69 (1H, s), 5.71 (1H,s), 5.03 (2H, d), 4.03 (3H, s), 3.84 (3H, s), 3.40 (8H, m), 2.70 (3H,s), 2.66 (3H, s); ³¹P NMR (CDCl₃): 6-10.01 (Ref=TPPO); HPLC (40%CH₃CN/0.1% TFA H₂O): 12.73 min; FAB MS: Calcd. for: 539.9898 found539.9904.

[0209] Scheme for Examples 14 and 15

EXAMPLE 14 2-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylN,N-bis(2-chloroethyl)phosphorodiamidate (6a)

[0210] n-Butyllithium (0.05 mL, 2.5M in hexanes) was added dropwise to asolution of 12 (25 mg, 0.11 mmol) in THF (3 mL. The solution was allowedto stir for 10 min at −78° C., and a solution of Cl₂P(O)N(CH₂CH₂Cl)₂ (34mg, 0.13 mmol) in THF (0.5 mL) was added all at once. The solution wasstirred for 1.5 h at −78° C. then warmed to −20° C. and ammonia gas wasbubbled through the reaction mixture for 7 min. The reaction was stirredfor an additional 8 min and then added to CH₂Cl₂/H₂O and extracted (3×).The combined organic layers were dried over Na₂SO₄ and evaporated.Column chromatography of the crude product (20% EtOAc/Acetone) afforded6a (27.6 mg, 59%) as a golden solid; R_(f)=0.47 (20% EtOAc/Acetone); ¹HNMR (CDCl₃): δ 6.68 (1H, s), 5.69 (1H, s), 5.01 (2H, m), 4.02 (3H, s),3.83 (3H, s), 3.65 (4H, m), 3.47 (4H, m), 2.03 (2H, bs); ³¹P NMR(CDCl₃): 6-9.35 (Ref=TPPO); HPLC (40% CH₃CN/0.1% TFA H₂O): 5.93 min; FABMS: Calcd. for: 446.0415 found 446.0419.

EXAMPLE 15 2-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylN,N-bis(2-bromoethyl)phosphorodiamidate (6b)

[0211] The title compound was prepared on a 0.11 mmol scale as describedabove for 6a, except that Cl₂P(O)N(CH₂CH₂Br)₂ was used as thephosphorylating agent. Column chromatography of the crude product (30%EtOAc/Acetone) afforded 6b (33.8 mg, 60%) as a peach solid; R_(f)=0.53(30% EtOAc/Acetone); ¹H NMR (CDCl₃): δ 6.69 (1H, s), 5.70 (1H, s), 5.05(2H, d), 4.02 (3H, s), 3.83 (3H, s), 3.473 (8H, m), 2.90 (2H, bs); ³¹PNMR (CDCl₃): 6-9.66 (Ref=TPPO); HPLC (40% CH₃CN/0.1% TFA H₂O): 7.02 min;FAB MS: Calcd. for: 511.9585 found 511.9579. Anal. Calcd. forC₁₅H₂₀Br₂N₃O₅P: C 35.11H 3.93 N 8.19 found C 35.33H 4.02 N 7.82.

EXAMPLE 16

[0212] The following illustrate representative pharmaceutical dosageforms, containing a compound of formula I (‘Compound X’), fortherapeutic or prophylactic use in humans. (i) Tablet 1 mg/tablet‘Compound X’ 100.0 Lactose 77.5 Povidone 15.0 Croscarmellose sodium 12.0Microcrystalline cellulose 92.5 Magnesium stearate 3.0 300.0 (ii) Tablet2 mg/tablet ‘Compound X’ 20.0 Microcrystalline cellulose 410.0 Starch50.0 Sodium starch glycolate 15.0 Magnesium stearate 5.0 500.0 (iii)Capsule mg/capsule ‘Compound X’ 10.0 Colloidal silicon dioxide 1.5Lactose 465.5 Pregelatinized starch 120.0 Magnesium stearate 3.0 600.0(iv) Injection 1 (1 mg/ml) mg/ml ‘Compound X’ (free acid form) 1.0Dibasic sodium phosphate 12.0 Monobasic sodium phosphate 0.7 Sodiumchloride 4.5 1.0 N Sodium hydroxide solution q.s. (pH adjustment to7.0-7.5) Water for injection q.s. ad 1 mL (v) Injection 2 (10 mg/ml)mg/ml ‘Compound X’ (free acid form) 10.0 Monobasic sodium phosphate 0.3Dibasic sodium phosphate 1.1 Polyethylene glycol 400 200.0 01 N Sodiumhydroxide solution q.s. (pH adjustment to 7.0-7.5) Water for injectionq.s. ad 1 mL

[0213] The above formulations may be obtained by conventional procedureswell known in the pharmaceutical art.

[0214] All publications, patents, and patent documents are incorporatedby reference herein, as though individually incorporated by reference.The invention has been described with reference to various specific andpreferred embodiments and techniques. However, it should be understoodthat many variations and modifications may be made while remainingwithin the spirit and scope of the invention.

What is claimed is:
 1. A compound a compound of formula I:

wherein: R¹ is an organic releasing group comprising a quinone ring;R_(a), R_(b), R_(c), and R_(d) are each independently hydrogen,(C₁-C₆)alkyl, or —CH₂CH₂X; and each X is independently halo,(C₁-C₆)alkylsulfonyl, halo(C₁-C₆)alkylsulfonyl, or arylsulfonyl, whereineach aryl is optionally substituted with one or more (e.g. 1, 2, 3, or4) halo, (C₁-C₆)alkyl, halo(C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)alkanoyl,(C₁-C₆)alkanoyloxy, (C₁-C₆)alkoxycarbonyl, cyano, nitro, ortrifluoromethoxy; provided at least two of R_(a), R_(b), R_(c), andR_(d) are —CH₂CH₂X; or a pharmaceutically acceptable salt thereof. 2.The compound of claim 1 wherein R¹ is a group of formula (II):

wherein R_(e) is hydrogen, halo, (C₁-C₆)alkyl, halo(C₁-C₆)alkyl,(C₁-C₆)alkoxy, (C₁-C₆)alkanoyloxy, cyano, nitro, or (C₁-C₆)alkylthio;and wherein the benz ring is optionally substituted by one or more (e.g.1, 2, 3, or 4) hydroxy, halo, (C₁-C₆)alkyl, halo(C₁-C₆)alkyl,(C₁-C₆)alkoxy, (C₁-C₆)alkylthio; (C₁-C₆)alkanoyl, (C₁-C₆)alkanoyloxy,(C₁-C₆)alkoxycarbonyl, cyano, nitro, mercapto, trifluoromethoxy, orNR_(f)R_(g); wherein each R_(f) and R_(g) is independently hydrogen,(C₁-C₆)alkyl, (C₁-C₆)alkanoyl, phenyl, benzyl, or phenethyl; or R_(f)and R_(g) together with the nitrogen to which they are attached arepyrrolidino, piperidino or morpholino.
 3. The compound of claim 1wherein R¹ is a group of formula (III):

wherein R_(k) is hydrogen or (C₁-C₆)alkyl; R_(m) is hydrogen or(C₁-C₆)alkyl, phenyl, benzyl, or phenethyl; and wherein the benz ring isoptionally substituted by one or two hydroxy, halo, (C₁-C₆)alkyl,halo(C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)alkylthio; (C₁-C₆)alkanoyl,(C₁-C₆)alkanoyloxy, (C₁-C₆)alkoxycarbonyl, cyano, nitro, mercapto,trifluoromethoxy, or NR_(f)R_(g); wherein each R_(f) and R_(g) isindependently hydrogen, (C₁-C₆)alkyl, (C₁-C₆)alkanoyl, phenyl, benzyl,or phenethyl; or R_(f) and R_(g) together with the nitrogen to whichthey are attached are pyrrolidino, piperidino or morpholino.
 4. Thecompound of claim 1 wherein R¹ is a group of formula (IV):

wherein R_(n) is hydrogen or (C₁-C₆)alkyl; R_(p) is hydrogen or(C₁-C₆)alkyl, phenyl, benzyl, or phenethyl; and wherein the benz ring isoptionally substituted by one or two hydroxy, halo, (C₁-C₆)alkyl,halo(C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)alkylthio; (C₁-C₆)alkanoyl,(C₁-C₆)alkanoyloxy, (C₁-C₆)alkoxycarbonyl, cyano, nitro, mercapto,trifluoromethoxy, or NR_(f)R_(g); wherein each R_(f) and R_(g) isindependently hydrogen, (C₁-C₆)alkyl, (C₁-C₆)alkanoyl, phenyl, benzyl,or phenethyl; or R_(f) and R_(g) together with the nitrogen to whichthey are attached are pyrrolidino, piperidino or morpholino.
 5. Thecompound of claim 1 wherein X is bromo, chloro, mesyl,trifluoromethylsulfonyl, or tosyl.
 6. The compound of claim 1 wherein Xis bromo.
 7. The compound of claim 2 wherein Re is hydrogen, halo,methyl, or methylthio.
 8. The compound of claim 3 wherein R_(k) ishydrogen or methyl.
 9. The compound of claim 3 wherein R_(m) is hydrogenor methyl.
 10. The compound of claim 4 wherein R_(n) is hydrogen ormethyl.
 11. The compound of claim 4 wherein R_(p) is hydrogen or methyl.12. The compound of claim 1 wherein R_(a) is (C₁-C₆)alkyl.
 13. Thecompound of claim 1 wherein R_(c) is (C₁-C₆)alkyl.
 14. The compound ofclaim 1 wherein R_(a) and R_(b) are each independently —CH₂CH₂X.
 15. Thecompound of claim 1 wherein R_(c), and R_(d) are each independently—CH₂CH₂X.
 16. The compound of claim 1 wherein R_(b) and R_(d) are eachindependently —CH₂CH₂X.
 17. The compound of claim 1 wherein R_(a) ismethyl.
 18. The compound of claim 1 wherein R_(c) is methyl.
 19. Thecompound of claim 1 wherein R_(a) and R_(b) are each —CH₂CH₂Br.
 20. Thecompound of claim 1 wherein R_(a), and R_(d) are each —CH₂CH₂Br.
 21. Thecompound of claim 1 wherein R_(b) and R_(d) are each —CH₂CH₂Br.
 22. Thecompound of claim 1 wherein R_(a) and R_(b) are each independently—CH₂CH₂Cl.
 23. The compound of claim 1 wherein R_(a), and R_(d) are eachindependently —CH₂CH₂Cl.
 24. The compound of claim 1 wherein R_(b) andR_(d) are each independently —CH₂CH₂Cl.
 25. The compound of claim 1which is: 2-(1,4-naphthoquinonyl)methyl N,N-bis(2-chloroethyl)phosphorodiamidate; 2-(3-Methyl-1,4-naphthoquinonyl)methylN,N-bis(2-chloroethyl) phosphorodiamidate;2-(3-Thiomethyl-1,4-naphthoquinonyl)methyl N,N-bis(2-chloroethyl)phosphorodiamidate; 2-(3-Bromo-1,4-naphthoquinonyl)methylN,N-bis(2-chloroethyl) phosphorodiamidate; 2-(1,4-Naphthoquinonyl)methylN,N-bis(2-bromoethyl) phosphorodiamidate;2-(3-Methyl-1,4-naphthoquinonyl)methyl N,N-bis(2-bromoethyl)phosphorodiamidate; 2-(1,4-Naphthoquinonyl)methyl bis[N-(2-chloroethyl)]phosphorodiamidate; 2-(1,4-Naphthoquinonyl)methylbis[N-methyl-N-(2-bromoethyl)]phosphorodiamidate;2-(3-Methyl-1,4-naphthoquinonyl)methyl bis[N-methyl-N-(2-bromoethyl)]phosphorodiamidate; 2-(1,4-Naphthoquinonyl)methylbis[N-methyl-N-(2-chloroethyl)] phosphorodiamidate;3-(5-Methoxy-1-methyl-4,7-indolequinonyl)-methylbis[N-methyl-N-(2-bromoethyl)] phosphorodiamidate;3-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylN,N-bis(2-bromoethyl)-phosphorodiamidate;2-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylbis[N-methyl-N-(2-bromoethyl)]phosphorodiamidate;2-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylN,N-bis(2-chloroethyl)-phosphorodiamidate; or2-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylN,N-bis(2-bromoethyl)-phosphorodiamidate; or a pharmaceuticallyacceptable salt thereof.
 26. The compound of claim 1 which is:3-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylN,N-bis(2-bromoethyl)-phosphorodiamidate;2-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylbis[N-methyl-N-(2-bromoethyl)]phosphorodiamidate;2-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylN,N-bis(2-chloroethyl)-phosphorodiamidate; or2-(5-Methoxy-1-methyl-4,7-indolequinonyl)methylN,N-bis(2-bromoethyl)-phosphorodiamidate; or a pharmaceuticallyacceptable salt thereof.
 27. A pharmaceutical composition comprising acompound of claim 1, in combination with a pharmaceutically acceptablediluent or carrier.
 28. A therapeutic method for preventing or treatingcancer comprising administering to a mammal in need of such therapy, aneffective amount of a compound of claim
 1. 29. The method of claim 28herein the cancer is a solid tumor.
 30. The method of claim 28 hereinthe cancer is a solid tumor.
 31. A method for preparing a compound offormula I as described in claim 1, wherein R¹ is a group of formula II,III, or IV, comprising oxidizing a corresponding compound of formula Iwherein R¹ is a group of formula VI, VII, or VIII.